Determinationofthetiter
Todeterminethetiter,anELISAwasperformedusingaserialdilutionoftheantibodydirectedagainstJarid1c(Cat.#25294).Theplateswerecoatedwiththepeptidesusedforimmunizationoftherabbit.Byplottingtheabsorbanceagainsttheantibodydilution(Figure1),thetiteroftheantibodywasestimatedtobe1:7,000.
WesternblotanalysisusingtheantibodydirectedagainstJarid1c
NuclearextractsofHeLacells(40µg)wereanalyzedbyWesternblotusingtheantibodyagainstJarid1c(Cat.#25294,lane2)andwiththepreimmuneserum(lane1)diluted1:1,000inTBS-Tweencontaining5%skimmedmilk.Thepositionoftheproteinofinterestisindicatedontheright(expectedsize:175kDa);theMarker(inkDa)isshownontheleft.Figure2showsthattheadditionalbandsatapproximately55and80kDaarealsopresentinthepreimmuneserum.
Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.
WB(1:1000)